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pgex 2t  (Addgene inc)


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    Addgene inc pgex 2t
    Pgex 2t, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pgextk+pak1/pGEXTK-Pak1+70-117+(Plasmid+%2312217)/bio_rxiv__64898__2026__02__24__707836-244-11-15
    Average 93 stars, based on 27 article reviews
    pgex 2t - by Bioz Stars, 2026-10
    93/100 stars

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    Plasmid Preparation:

    Article Title: Crystal structure of a novel guanine nucleotide exchange factor encoded by the scrub typhus pathogen Orientia tsutsugamushi
    Article Snippet: .. The protein was diluted to 40 mg/ml, supplemented with MgCl 2 to 5 mM final, flash frozen in 20 μl aliquots and stored at - 80°C. pGEXTK-Pak1 70-117 was a gift from Jonathan Chernoff (Addgene plasmid # 12217; http://n2t.net/addgene:12217 ; RRID:Addgene_12217) ..

    Article Title: Protease activated receptor-1 (PAR1) derived cytoprotective polypeptides and related methods
    Article Snippet: .. The pGEXTK-PAK1 70-117 construct encoding a GST fusion to p21-activated kinase (PAK-1)-binding domain (PBD) was kindly made available by Dr. J Chernoff (Addgene plasmid 12217). .. GST-PAK1 was purified from transformed BL21 (DE3) Escherichia coli using B-Per lysis buffer (Pierce) with lysozyme, DNAse 1 and Halt EDTA-free protease inhibitor cocktail (Pierce) on Glutathione-agarose according to the manufacturer's recommendation.

    Article Title: Protease Activated Receptor-1 (PAR1) Derived Cytoprotective Polypeptides and Related Methods
    Article Snippet: .. The pGEXTK-PAK1 70-117 construct encoding a GST fusion to p21-activated kinase (PAK-1)-binding domain (PBD) was kindly made available by Dr. J Chernoff (Addgene plasmid 12217). .. GST-PAK1 was purified from transformed BL21 (DE3) Escherichia coli using B-Per lysis buffer (Pierce) with lysozyme, DNAse I and Halt EDTA-free protease inhibitor cocktail (Pierce) on Glutathione-agarose according to the manufacturer's recommendation.

    Construct:

    Article Title: Protease activated receptor-1 (PAR1) derived cytoprotective polypeptides and related methods
    Article Snippet: .. The pGEXTK-PAK1 70-117 construct encoding a GST fusion to p21-activated kinase (PAK-1)-binding domain (PBD) was kindly made available by Dr. J Chernoff (Addgene plasmid 12217). .. GST-PAK1 was purified from transformed BL21 (DE3) Escherichia coli using B-Per lysis buffer (Pierce) with lysozyme, DNAse 1 and Halt EDTA-free protease inhibitor cocktail (Pierce) on Glutathione-agarose according to the manufacturer's recommendation.

    Article Title: Protease Activated Receptor-1 (PAR1) Derived Cytoprotective Polypeptides and Related Methods
    Article Snippet: .. The pGEXTK-PAK1 70-117 construct encoding a GST fusion to p21-activated kinase (PAK-1)-binding domain (PBD) was kindly made available by Dr. J Chernoff (Addgene plasmid 12217). .. GST-PAK1 was purified from transformed BL21 (DE3) Escherichia coli using B-Per lysis buffer (Pierce) with lysozyme, DNAse I and Halt EDTA-free protease inhibitor cocktail (Pierce) on Glutathione-agarose according to the manufacturer's recommendation.

    other:

    Article Title: Geranylgeranyl diphosphate synthase inhibition induces apoptosis that is dependent upon GGPP depletion, ERK phosphorylation and caspase activation
    Article Snippet: The anti-tubulin (E7) antibody developed by Michael Klymkowsky was obtained from the Developmental Studies Hybridoma Bank at the University of Iowa. pGEXTK-Pak1 70-117 was obtained from Addgene (Cambridge, MA, USA).



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    Circulating levels of norepinephrine and angiotensin II are elevated during chronic hypertrophic stress. Activation of the β 1 AR elicits G s -dependent activation of adenylyl cyclase, leading to cAMP production and protein kinase A (PKA) activation. Active PKA phosphorylates substrates critical for excitation-contraction coupling. AT1R activation leads to Rac1 activation and promotes Rac1 S -palmitoylation cycling. S -palmitoylated Rac1 is required to evoke dephosphorylation of PKA substrates and dampen excessive adrenergic signaling that promotes cardiac maladaptation, likely through <t>Pak1-dependent</t> activation of PP2A. Upon loss of Rac1 palmitoylation cycling at Cys-178 (as observed in Rac1 cKI hearts), PP2A-mediated antagonism of PKA substrate activity is not properly regulated, resulting in hyperphosphorylation of PKA substrates that chronically promotes cardiac decompensation, systolic dysfunction, and adverse hypertrophic remodeling. AC, adenylyl cyclase; AngII, angiotensin II; AT1R, angiotensin II receptor type I; β 1 AR, β1 adrenergic receptor; LTCC, L-type calcium channel; MyBPC3, myosin binding protein C3; NE, norepinephrine; Pak1, p21-activated kinase-1; PKA, protein kinase A; PLN, phospholamban; PP2A, protein phosphatase 2A; SERCA, sarco-endoplasmic reticulum calcium ATPase; TnI, troponin I; TnT, troponin T.
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    Circulating levels of norepinephrine and angiotensin II are elevated during chronic hypertrophic stress. Activation of the β 1 AR elicits G s -dependent activation of adenylyl cyclase, leading to cAMP production and protein kinase A (PKA) activation. Active PKA phosphorylates substrates critical for excitation-contraction coupling. AT1R activation leads to Rac1 activation and promotes Rac1 S -palmitoylation cycling. S -palmitoylated Rac1 is required to evoke dephosphorylation of PKA substrates and dampen excessive adrenergic signaling that promotes cardiac maladaptation, likely through Pak1-dependent activation of PP2A. Upon loss of Rac1 palmitoylation cycling at Cys-178 (as observed in Rac1 cKI hearts), PP2A-mediated antagonism of PKA substrate activity is not properly regulated, resulting in hyperphosphorylation of PKA substrates that chronically promotes cardiac decompensation, systolic dysfunction, and adverse hypertrophic remodeling. AC, adenylyl cyclase; AngII, angiotensin II; AT1R, angiotensin II receptor type I; β 1 AR, β1 adrenergic receptor; LTCC, L-type calcium channel; MyBPC3, myosin binding protein C3; NE, norepinephrine; Pak1, p21-activated kinase-1; PKA, protein kinase A; PLN, phospholamban; PP2A, protein phosphatase 2A; SERCA, sarco-endoplasmic reticulum calcium ATPase; TnI, troponin I; TnT, troponin T.

    Journal: JCI Insight

    Article Title: Rac1 palmitoylation is required for cardiac stress adaptation and regulation of protein kinase A signaling

    doi: 10.1172/jci.insight.193733

    Figure Lengend Snippet: Circulating levels of norepinephrine and angiotensin II are elevated during chronic hypertrophic stress. Activation of the β 1 AR elicits G s -dependent activation of adenylyl cyclase, leading to cAMP production and protein kinase A (PKA) activation. Active PKA phosphorylates substrates critical for excitation-contraction coupling. AT1R activation leads to Rac1 activation and promotes Rac1 S -palmitoylation cycling. S -palmitoylated Rac1 is required to evoke dephosphorylation of PKA substrates and dampen excessive adrenergic signaling that promotes cardiac maladaptation, likely through Pak1-dependent activation of PP2A. Upon loss of Rac1 palmitoylation cycling at Cys-178 (as observed in Rac1 cKI hearts), PP2A-mediated antagonism of PKA substrate activity is not properly regulated, resulting in hyperphosphorylation of PKA substrates that chronically promotes cardiac decompensation, systolic dysfunction, and adverse hypertrophic remodeling. AC, adenylyl cyclase; AngII, angiotensin II; AT1R, angiotensin II receptor type I; β 1 AR, β1 adrenergic receptor; LTCC, L-type calcium channel; MyBPC3, myosin binding protein C3; NE, norepinephrine; Pak1, p21-activated kinase-1; PKA, protein kinase A; PLN, phospholamban; PP2A, protein phosphatase 2A; SERCA, sarco-endoplasmic reticulum calcium ATPase; TnI, troponin I; TnT, troponin T.

    Article Snippet: For experiments in NRCMs with overexpressed Rac1, we generated a recombinant glutathione S -transferase (GST)-PAK-PBD fusion protein by transforming BL21-DE3 E . coli with a pGEXT-PAK-PBD-70-117 plasmid that contains residues 70–117 of human Pak1 (Addgene, 12217, gift from Jonathan Chernoff).

    Techniques: Activation Assay, De-Phosphorylation Assay, Activity Assay, Binding Assay